CyDye DIGE fluors are available as minimal and saturation labeling dyes.
- Allow detection of up to three prelabelled protein samples and standards on the same 2-D electrophoresis gel
- Size- and charge-matched dyes enable co-migration of labelled samples within the gel
- Bright and highly sensitive dyes allow the use of the minimal labelling technique
- Minimal loss of signal during labelling, separation, and scanning
- No change in signal over wide pH range used during first-dimension (IEF) separation
- Discrete signal from each fluor with minimal cross-talk contributes to high accuracy
- Soluble in cold and hot water (solid)
CyDye DIGE fluors are exceptional dyes for multicolor analysis, offering bright and intense colors with narrow excitation and emission bands. The fluors are spectrally distinct, making them highly suitable for multicolor detection. CyDye DIGE fluors utilize these benefits but are also size- and charge-matched for 2-D DIGE using Ettan DIGE system.
CyDye DIGE fluor labeling kits for scarce samples:
Protein samples and the internal standard are each labeled with one CyDye DIGE Fluor minimal dye. These labeled samples are then combined, run on an isoelectric focusing gel in the first dimension, and separated by SDS-PAGE in the second dimension. Electrophoresis is simplified with Ettan IPGphor 3, or Multiphor II with Immobiline DryStrip gels in the first dimension and Ettan DALTtwelve or Ettan DALTsix electrophoresis systems in the second dimension.
The NHS ester reactive group of CyDye DIGE Fluor minimal dyes covalently attaches to the epsilon amino group of lysine of proteins via an amide linkage. The ratio of dye to protein has been designed to ensure the dyes are limiting in the reaction. As a result, approximately 3% of the available proteins are labeled and then only on a single lysine per protein (one dye per protein, or minimal labeling).
The amino acid lysine in proteins carries an intrinsic single positive charge at neutral or acidic pH. CyDye DIGE Fluor minimal dyes also carry a single positive charge which, when coupled to the lysine, replaces the single positive charge of the lysine with its own, ensuring that the pI of the protein is not significantly altered when compared with the same unlabeled protein.
The minimal dyes are intended for general 2-D application use where sufficient amounts of sample are available. The saturation dyes from the scarce sample labeling kit are designed to be used for applications where only small amounts of sample are available, for example in laser capture microdissection.
Ordering information: Dimethylformamide (DMF) has to be ordered separately.
Caution: Estimated shelf life from manufacture date: 8 - 12 months, for unreconstituted dye.
